Hemoglobin solution and method



Patented Oct. 24, 1950 a UNITED nEMoGLoBrN sr rEs PATENT OFFICE John 0.Bower, Wyncote, Pa.

NoDrawing.

Application January 25, 1944, Serial No. 519,686

' P6 Claims. (01.106-161) My invention relates to hemoglobin solutions,that is to say, solutions comprised of the content of human blood cells,that substancecontained within the blood cell Walls. i

This application is a continuation-impart of my application Serial No.502,503, filed September 15, 1943, now Patent No. 2,493,943. 7

The prime object of my invention is to obtain an hemoglobin solutionuncontaminated at any time in its preparation by matter foreign to theblood itself.

In the past hemoglobin solutionshave been,

matter foreign to the blood and the use of any of them.

A further object of my invention is the effecting of the solution underconditions which preclude to a very maximum degree contaminationeitherfrom atmosphere or from handling.

The objects of my invention I [attain .outstandingly by mechanicallyinstead of chemically dissolving the cell walls. I comprise the solutionof hemoglobin containing blood cell walls inechanically dissolved tosubmicroscopic fineness.

The method I use consists of freezing at such an extreme rate and atsuch a lowdegree that the cell walls are submicroscopically dividedyre--duced to a state of complete structural dissolution, and enter intosolution with the liquid con: tent of the blood when the frozen mixtureis thawed. The thawed such solution is the product of my invention.

The method and the product are thus intimately inter-related.

More specifically, my method is the following:

Blood is drawn from a donor by the closed method, a well-known method inwhich there are no contacts of the blood with outside air. It is drawninto a donor bottle either with or with out sodium citrate, the usualintake tubing, needle, outlet tubing and cotton filter being em ployed.As soon thereafter as possible this blood, is centrifuged and thesupernatent plasma, or serum withdrawn, this centrifuging and witha issimilar to a drawal likewise being by the closed method. The red cellresidue is that which I use. It may be used at any time after plasmaextraction before it becomes spoiled. It will keep Well in atmosphere atroom temperature for some days, and will keep several weeks in arefrigerator.

I pour this residue into a suitable container solid. carbon marketed bythe Sun Oil Company, and highly refined kerosene in its properties.) Thetemperature to which the red cell mixture is frozen and the rate of suchfreezing are those which produce the same results as are obtained byimmersing for from two to three minutes a 30 cc. glass test tube filledwith red cell mixture in a bath of bath the solid frozen material isdark pink in color rather than red, and the surface of the solid is inthe center elevated above its original surface level. In a 30 cc. testtube treatment the elevation is in the form of a nipple-like projection.'If the red blood cell mixture contains a substantial percentage ofplasma, it will be observed in this elevated portion of the mixture, andthe elevated portion will be somewhat lighter in color than the mainbody. The mixture so frozen is removed from the solution at the com-.pletion of its freezing and is permitted to thaw (preferably a gradualthawing to prevent damage to containing vessels), whereupon one has theproduct of my inventionthe hemoglobin solution comprised of the proteinand other content of blood cells and th mechanically dissolved cellwalls.

In the product of my invention I believe the mechanically dissolved cellwalls are carried in a state of suspension. The solution appears to Bymy method the dissolution of be colloidal. the cell walls is achieved tosuch a high degree of fineness that the suspended cannot bedifferentiated from matter with microscopes of though perhaps they canbe the hemoglobin ordinary power,

(Sunoco spirits is a petroleum hydrocell wall matter differentiated byusing microscopes of high power. In referring to microscopes of ordinarypower, I have in mind the conventional laboratory instrument, which iscapable of producing magnifications of from 100 diameters up to 600 oreven 900 diameters. How- 5 ever. the degree of fineness of thedissolution of the cell walls may vary without departing from myinvention, either its method phase or its product phase. I can, forexample,lregulate the'g degree of fineness of the dissolution of thesub-' microscopic dimensions by changing the tem; perature at which thecell walls the rate of such freezing. L

This product of my invention has many uses, some of which are more fullydeveloped in other uses are intravenous injection into the human bloodstream; application as a solution to both clean and aseptic wounds; thecoating or irffpregnating of bone, tissue, or other filling as appliedto a wound; the coating orimpr'egnatin'g of sutures of everydescription, particularly sutures composed of animal fiber such as the"well-known .catg'ut sutures; and ;a synthetic' spinning of the mainbodies of sutures fr m my hemoglobin solution itself; I When injected.intothe human blood stream it-affords the blood making processes of thef human body the-materials with which .to make fresh bloodand replenishthe blood supply and' so ,nourish and strengthen the body. For injec'-'tion the solution is brought by plasma additions to the sameconcentration as the blood ofth'e patient. It is my belief that thecomplete de'=' the type of the originalblood from which-thesolution wasderivedand renders the solutioncompletely adaptable to the type of bloodof the} individual subjected to the injection. From this standpoint aninjection practice utilizing my solution has a markedadvantageover andabove ordinary. transfusion. practices which are handicapped inadaptability by complete subjec tion to type. 1 Inall following uses thesolution constitutes a wound healing product of great" value in" medicine. Outstandingly, this isbecause it contains practically all. of theconstituents used by: the" humanbody in itsprocesses of-healingi -It-ctains themin a state more nearly approachin the ideal state than anyother such solution-thus fardiscovered. Moreover, these constituent rein a state of substantially perfect purity, free alike from bacteria andirritating-or retard foreign substances. This is by reason, firs t,:that the closed methods of deriving the red blood c'ell mixture and thefollowing freezing at=extreme sub-zero temperatures respectively preventen- I trance of bacteria and inhibitbacteria growth; and, second, thatno substance whatever other than the blood substance is introduced tothe mixture or the original solution. The chemicals of the old-fashionedsolutions are avoided altogether. 1 7 As applied .locally'to eitherclean orunclean wounds either before or after the making or occurrenceof thevwound, the solution'is poured, painted, or otherwise spread .onin airelatively ii viscous state. It Jdries rapidly; presentingl'iasmooth, varnish-like superior-surface *Several applications may. be madetop'roduce a ccv'ering or filling of any desired thickness.Thevisco'sit; may; be controlled by. adding blood plasma to reduce it,orztakin outplasma and water con tent toa increaseit. Thezwater:contentiofthe copending applications. Prominent among such air red bloodcell residue from plasma production from human blood is to and theplasma content but a few percent, the great bulk being removed by thecentrifuging process of plasma extraction. For ordinary shallow woundssuch as cuts and abrasions, a very light applied coating quickly forms askin which so protects the wound as to render necessary but a very lightdressing. It penetrates the epidermis without irritation 'afid'stronglyadheres to it, effectively ,seallng the sides of any superficial woundto the are frozen and i .cially valuablesubs'titute for iodine inpreparadjoining skin. This quality makes it an espeing the epidermisareas adjoining the location of an intended incision, and guarding thewound against infection. 1'; av coating for fillings or applications ofbone tissue or other wound-filling substance to be grb'wn to or into thebody, it provides directly in the joint between the applied material orfilling the substance of which the joint itselfis ultimately'to be madeby the body processes. The body processes absorb this material withoutthe slightest irritation to the wound or the adjoin irig bodyparts.Applications of the solution to these'ends can be made by pouring orpainting upon the open wound, precoating the application or filling'bydipping, soaking, painting",- or otherwise placing upon or within thewound in' j'ecting the solution'between the filling or application andthe wound by suitable injection means, orby any other convenient means.

In the trea'tmentof existing sutures of fiber or other such material, Iusually im'rner'se struction of the blood cells destroys altogether 3bwhether I 'de'sire the suture to be thoroughly impregnated. I have usedall of these methods on different kinds of sutures, notably upon'catg'ut sutures but also sutures of silk, of casein, I have appliedcoatings and of other materials. of different thicknesses andimpregnations of different degree. I have subsequently embeddedthesesutures in the stomach walls of rabbits and taken microscopic sectionsat different'periods of the healing. In all cases I have found highlybeneficial results The healing in the case of :coated 'or impregnatedcatgut sutures occurs in a minimumtime and with minimal irritation tothe walls through which the suture passes.

absorption is rapid and the'time only a minor fraction of that requiredwhen ordinary such sutures are-similarly used. v I pregnated sutures ofsilk, while untreated such sutures are relatively unabsorbable, theindications are that the impregnated silk suture is rendered absorbable,in whole or in part, and in ,any case that they produce a veritableminimum of irritation.

Finally, I have synthetically spun sutures this solution and embeddedthese synthetically spun sutures in the stomach walls of rabbits andfound them still more readily absorbed thanthe solution impregnatedcatgut (but yet taking amply adequate absorption time to permit properhealing) and thorough healing process. In synthetically spin-Q wi hingthis solution I modify it by introducing sodium sulphide to carry thecell walls already in solution by suspension as well intochemicalsolution. I introduce to my original hemoglobinsolution abovedescribed a 20% solution of'sodium qssfisulphideinthe-proportion of'onepart by volume In the case of finaccompanied by a still more toapproximately three parts of the original hemoglobin solution in whichthe cell walls are mechanically dissolved. This I introduce through asuitable spinning nozzle into a suitable precipitant, forming a sutureof adequate cross-section. This precipitated suture I then introduce toa setting bath following a general order of steps such as very commonlyused in spinning synthetic filaments.

Obviously both the product of my invention (the solution) and the methodof producing it, intimately related though they are, are each subjecttomodification without in any wise departing from the generic spirit ofmy invention. For example, animal blood cells other than human bloodcells may be used in certain cases. So also are the various and sundryutilizations of it. Likewise the manner of handling it may be variedwithout departing in any wise from the generic spirit.

Speaking of the manner of handling, it is to be remarked that one of theoutstanding merits of my invention is the aseptic character of thesolution. It will keep fresh and free from deterioration,bacteriologically or otherwise, for long periods of time. It possessesnaturally a very high resistance to deterioration, and aided by knownprocesses of refrigeration and other protections may be keptindefinitely. Coated or impregnated sutures keep well either wet or dry.Wet they may be preserved in xylol as readily as any other suture. Dry,either as coated or impregnated, the superficial coating is tenaciouslyadherent and presents a smooth, ibright, protective surface free fromcracking when bent, like a pliable varnish or enamel. 7

Irrespective of the circumstantial nature of the terminology of theannexed claims, the intent of this specification and claims is toprotect to myself my invention in all the fullness of its genericspirit. For example, so long as the primary characteristics of thesolution are not substantially impaired thereby, various additions to itmay be made. The expression consisting essentially, in the claims whichfollow, is not to be construed as excluding the presence of suchadditions.

What is claimed is:

1. The method of preparing an hemoglobin solution comprising blood cellstructure, which method consists of freezing the blood cells to atemperature and at a rate of freezing corresponding to that resultingfrom the immersion of a 30 cc. test tube containing blood cells in afreezing solution at approximately 85 F. for from two to three minutes.

2. The liquid of claim 5, in which the sodium sulfide constitutes about5% of the total volume.

3. The method of preparing an hemoglobin spinning solution comprisingthe elements of blood cell structure, which method comprises freezingand thawing the blood cells at a rate and to a temperature yielding thesame results as follow the immersion of a 30 cc. test tube filled withred cell mixture in a bath of Sunoco Spirits and dry ice atsubstantially F. for from 2 to 3 minutes, thawing the mixture, andadding an aqueous solution of sodium sulfide.

4. The method of claim 3, in which the sodium sulfide is added inproportions of about one part of a 20% aqueous solution thereof to aboutthree partsof blood cell mixture.

5. A spinnable liquid consisting essentially of sodium sulfide and aconstituency of blood which has been frozen at a rate and'to atemperature yielding the same results as follow the immersion of a 30cc. test tube filled with red cell mixture in a bath of Sunoco spiritsand dry ice at substantially -85 F. for from two to three minutes, andsubsequently thawed.

6. A solution consisting of a frozen and thawed material selected fromthe group which consists of who1e blood, whole blood from which serumhas been separated, citrated whole blood, and citrated whole blood fromwhich plasma has been separated, which solution has the elements ofstroma in a state of dissolution in blood cell content and hasthroughout the clarity and filtering characteristics which result uponthe freezing of red blood cell mixture in a 30 cc. test tube in afreezing solution at about -85 F. for from 2 to 3 minutes, followed bythawing of such frozen mixture.

JOHN O. BOWER.

REFERENCES CITED The following references are of record in the file ofthis patent:

UNITED STATES PATENTS Number Name Date 1,380,427 Sgalitzer June 7, 19212,287,028 DAmbrosio et a1. June 23, 1942 FOREIGN PATENTS Number CountryDate 165,832 Great Britain July 11, 1921 422,990 Great Britain June 23,1935 6,700 Great Britain of 1898 OTHER REFERENCES Sci. Amen, Mar.

Certificate of Correction Patent N 0. 2,527,210 October 24, 1950 JOHN0'. BOWER It is hereby certified that error appears in the printedspecification of the above numbered patent requiring correction asfollows:

Column 6, list of references cited, line 47, for June 23, 1935 readJanuary 23, 1935; line 51, for (1932) read (1923); line 59, for (1939)read (1938); and that the said Letters Patent should be read ascorrected above, so that the same may conform to the record of the casein the Patent Oifice.

Signed and sealed this 20th day of February, A. D. 1951.

THOMAS F. MURPHY,

Assistant Commissioner of Patents.

1. THE METHOD OF PREPARING AN HEMOGLOBIN SOLUTION COMPRISING BLOOD CELLSTRUCTURE, WHICH METHOD CONSISTS OF FREEZING THE BLOOD CELLS TO ATEMPERATURE AND AT A RATE OF FREEZING CORRESPONDING TO THAT RESULTINGFROM THE IMMERSION OF A 30 CC. TEST TUBE CONTAINING BLOOD CELLS IN AFREEZING SOLUTION AT APPROXIMATELY -85*F. FOR FROM TWO TO THREE MINUTES.